[NMR paper] NMR Spectroscopic Studies Reveal the Critical Role of Isopeptide Bond in Forming the Otherwise Unstable SpyTag-SpyCatcher Mutant Complexes.
Related ArticlesNMR Spectroscopic Studies Reveal the Critical Role of Isopeptide Bond in Forming the Otherwise Unstable SpyTag-SpyCatcher Mutant Complexes.
Biochemistry. 2020 May 29;:
Authors: Zhang N, Liu J, Liu Y, Wu W, Fang J, Da XD, Wang S, Zhang WB
Abstract
The interplay between protein folding and chemical reaction has been an intriguing subject. In this contribution, we report the study of SpyTag and SpyCatcher reactive mutants using combined techniques of SDS-PAGE, LC-MS, circular dichroism, and NMR spectroscopy. It was found that the wild-type SpyCatcher is well-folded in solution and docks with SpyTag to form an intermediate that promotes isopeptide formation. By contrast, the double mutant SpyCatcherVA is disordered in solution, yet remains reactive toward SpyTag, forming a well-folded covalent complex. Control experiments using the catalytically inactive mutants further reveal the critical role of the isopeptide bond in stabilizing the otherwise loose SpyTag-SpyCatcherVA complex, amplifying the effect of minute sequence disparity. We believe that the synergy between protein folding and isopeptide bonding is an effective way to enhance protein stability and engineer protein-protein interactions.
PMID: 32469203 [PubMed - as supplied by publisher]
[NMR paper] Biophysical studies and NMR structure of YAP2 WW domain - LATS1 PPxY motif complexes reveal the basis of their interaction.
Biophysical studies and NMR structure of YAP2 WW domain - LATS1 PPxY motif complexes reveal the basis of their interaction.
Biophysical studies and NMR structure of YAP2 WW domain - LATS1 PPxY motif complexes reveal the basis of their interaction.
Oncotarget. 2018 Jan 30;9(8):8068-8080
Authors: Verma A, Jing-Song F, Finch-Edmondson ML, Velazquez-Campoy A, Balasegaran S, Sudol M, Sivaraman J
Abstract
YES-associated protein (YAP) is a major effector protein of the Hippo tumor suppressor pathway, and is phosphorylated by the...
nmrlearner
Journal club
0
03-01-2018 09:20 PM
A non-uniform sampling approach enables studies of dilute and unstable proteins
A non-uniform sampling approach enables studies of dilute and unstable proteins
Abstract
NMR spectroscopy is a powerful method in structural and functional analysis of macromolecules and has become particularly prevalent in studies of protein structure, function and dynamics. Unique to NMR spectroscopy is the relatively low constraints on sample preparation and the high level of control of sample conditions. Proteins can be studied in a wide range of buffer conditions, e.g. different pHs and variable temperatures, allowing studies of proteins under...
nmrlearner
Journal club
0
02-11-2017 12:49 PM
[NMR paper] Quantitative Analysis of STD-NMR Spectra of Reversibly Forming Ligand-Receptor Complexes.
Quantitative Analysis of STD-NMR Spectra of Reversibly Forming Ligand-Receptor Complexes.
Related Articles Quantitative Analysis of STD-NMR Spectra of Reversibly Forming Ligand-Receptor Complexes.
Top Curr Chem. 2008;273:15-54
Authors: Krishna NR, Jayalakshmi V
Abstract
We describe our work on the quantitative analysis of STD-NMR spectra of reversibly forming ligand-receptorcomplexes. This analysis is based on the theory of complete relaxation and conformational exchange matrixanalysis of saturation transfer (CORCEMA-ST) effects. As part...
nmrlearner
Journal club
0
04-24-2013 09:48 PM
NMR spectroscopic and theoretical analysis of a spontaneously formed Lys-Asp isopeptide bond.
NMR spectroscopic and theoretical analysis of a spontaneously formed Lys-Asp isopeptide bond.
NMR spectroscopic and theoretical analysis of a spontaneously formed Lys-Asp isopeptide bond.
Angew Chem Int Ed Engl. 2010 Nov 2;49(45):8421-5
Authors: Hagan RM, Björnsson R, McMahon SA, Schomburg B, Braithwaite V, Bühl M, Naismith JH, Schwarz-Linek U
nmrlearner
Journal club
0
02-15-2011 07:17 PM
[NMR paper] The role of disulfide bond in the amyloidogenic state of beta(2)-microglobulin studie
The role of disulfide bond in the amyloidogenic state of beta(2)-microglobulin studied by heteronuclear NMR.
Related Articles The role of disulfide bond in the amyloidogenic state of beta(2)-microglobulin studied by heteronuclear NMR.
Protein Sci. 2002 Sep;11(9):2218-29
Authors: Katou H, Kanno T, Hoshino M, Hagihara Y, Tanaka H, Kawai T, Hasegawa K, Naiki H, Goto Y
beta(2)-Microglobulin (beta2-m) is a major component of dialysis-related amyloid fibrils. Although recombinant beta2-m forms needle-like fibrils by in vitro extension reaction at pH...
nmrlearner
Journal club
0
11-24-2010 08:58 PM
[NMR paper] Real-time and equilibrium (19)F-NMR studies reveal the role of domain-domain interact
Real-time and equilibrium (19)F-NMR studies reveal the role of domain-domain interactions in the folding of the chaperone PapD.
Related Articles Real-time and equilibrium (19)F-NMR studies reveal the role of domain-domain interactions in the folding of the chaperone PapD.
Proc Natl Acad Sci U S A. 2002 Jan 22;99(2):709-14
Authors: Bann JG, Pinkner J, Hultgren SJ, Frieden C
PapD is a periplasmic chaperone essential for P pilus formation in pyelonephritic strains of E. coli. It is composed of two domains, each of which contains a tryptophan...
NMR Spectroscopic Studies Reveal the Critical Role of Isopeptide Bond in Forming the Otherwise Unstable SpyTag-SpyCatcher Mutant Complexes.
Linear Mode
